PEPTIDE FOR USE IN SIMULTANEOUS PROTEIN QUANTIFICATION OF METABOLIZING ENZYMES USING MASS SPECTROMETRIC ANALYSIS APPARATUS

摘要

There are provided a peptide consisting of an amino acid sequence for simultaneously quantifying absolute amounts of metabolizing enzyme proteins in a biological sample at high sensitivity and a method for using the same. A peptide which can be detected at high sensitivity with a mass spectrometer that enables highly sensitive simultaneous quantification of metabolizing enzymes, intracellular proteins, is selected, and the amino acid sequence thereof is identified. Using a stable-isotope-labeled peptide having the same amino acid sequence as the amino acid sequence of this peptide to be quantified, a mass spectrometry by LC-MS/MS is performed at predetermined concentration levels of the stable-isotope-labeled peptide to create a calibration curve. The stable-isotope-labeled peptide is added to a peptide fragment obtained by fragmenting metabolizing enzyme proteins to be quantified in a sample with trypsin, a mass spectrometry by LC-MS/MS is performed to calculate a mass spectrum area ratio of the metabolizing enzyme protein peptides to be quantified and the stable-isotope-labeled peptide, and a quantitative value is obtained from the area ratio using the calibration curve.